As. J. Food Ag-Ind.

2008, 1(02), 97-104

Asian Journal of
Food and Agro-Industry
ISSN 1906-3040
Available online at www.ajofai.info
Research Paper

Effects of soy protein isolate on chemical and physical

characteristics of meat analog
Kajirat Rareunrom, Sunanta Tongta* and Jirawat Yongsawatdigul
School of Food Technology, Institute of Agricultural Technology, Suranaree University of
Technology, Nakhon Ratchasima 30000, Thailand
*Author to whom correspondence should be addressed, email:
Paper originally presented at Food Innovation Asia 2007
_______________________________________________________________________________

Abstract: Soy protein analog was fabricated using a twin screw extruder. Different soy
protein isolate (SPI) contents of 20, 40, 60 and 80% were blended with defatted soy flour
(DSF) and then fed into an extruder with the feed rate of 65 g/min. The barrel temperature
profile was 60, 90,140 and 160oC. The screw speed was 250 rpm. Based on protein
solubility studies using different buffer solutions containing phosphate, sodium dodecyl
sulphate, urea or 2-mercaptoethanol, disulfide bond, hydrophobic interaction and
hydrogen bond were major linkages stabilizing meat analog structure. Increasing SPI
content from 20 to 80% did not increase these chemical linkages but resulted in a greater
expansion ratio (p<0.05). Cutting force, which was monitored by a texture analyzer,
decreased with increasing SPI content. Textural characteristics, as judged by tearing, and
fibrous characteristics using Quantitative Descriptive Analysis were reduced with
increased SPI contents.
Keywords: food processing, meat analog, soy protein isolate, chemical linkages, texture,
twin screw extruder
________________________________________________________________________
Introduction
Meat analog is a major type of texturized plant protein (TPP) which is extensively used to
imitate meat products (Sheard, Ledward and Mitchell, 1984). It is considered to be a
health food which contains high protein and fibre, particularly suited to vegetarian diets.
The other beneficial characteristics of TPP are year-long shelf stability in dry form and
capacity for rehydration without loss in structure, shape and chewy texture (Harper, 1981).
Soy protein is a major important component to fabricate the structure of meat analog. The
value of soy protein is comparable to that of meat protein (deMan, 1990). Furthermore, the
risk of heart disease and cancer of consumers is also reduced (Liu, 1997). Extrusion
cooking is the principle processing method used to fabricate meat like texture and

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98

plexilamellar structure for soy protein products (Burgess and Stanley, 1976). It also offers
some advantages in terms of cost and the capability for high volume production (Harper,
1981).
The structure of texturized soy protein (TSP) products depends on production conditions
and material components (Ha, 1995; Sheard et al., 1984). Quantity and quality of soy
protein are the important factors that affect the chemical and physical characteristics of
meat analog. Maurice and Stanley (1978) suggested that increasing protein content
resulted in a high shearing force on the fine texture of extrudate. Sheard and others (1984)
reported that soy carbohydrate also supported the main structure of soy protein extrudate.
The fibre and layer structures of meat analog were generated by protein texturization
during extrusion. Thermal denaturation set the protein matrix in a fibre-like configuration
involving protein cross-linking and interaction between protein and other components
during extrusion. Hager (1984) mentioned that chemical bonds, which were disulfide
bonds, hydrogen bonds and hydrophobic interaction, were present to support the stabilized
extrudate structure and new chemical linkages during extrusion cooking did not appear.
However, Hayakawa (1992) reported that the disulfide bond was disrupted during
processing, but did not affect the extrudate structure. The objectives of this study were to
investigate the chemical linkages and physical characteristics of soy protein meat analog
containing different soy protein isolate contents.
Materials & Methods
Materials
Defatted soy flour (DSF), soy protein isolate (SPI, Profam 970) were supplied by ADM
protein specialties, Decatur IL, USA. DSF contained 50% protein and 0.22% fat. The
protein content of SPI was 90%. The protein and fat content of materials were determined
using the method of AOAC (1995). Urea was obtained from Sigma-Aldrich Gmblt,
Germany. The other reagents were of analytical grade.
Extrusion processing
DSF was blended with SPI in order to obtain 20, 40, 60 and 80% SPI. All materials were
extruded using an APV Baker MPF 19/25 twin screw extruder (APV Baker, UK) which
was equipped with a 3 mm diameter die. The dry feed rate and screw speed were kept
constant at 65 g/min and 250 rpm respectively. Water was added to the blends to obtain
30% moisture content. Barrel temperature profile was set at 60, 90, 140 and 160C from
zone 1 to zone 4, respectively. After the product was extruded from the die, it was dried in
a hot air oven at 50C for 3 hrs.
Protein solubility determination
Protein solubility of both raw materials and extrudates were analyzed using the modified
method of Lin et al. (2000). Buffer solution used for protein extraction included; (1)
0.035M phosphate buffer (pH 7.6), (2) 1.5% sodium dodecyl sulfate (SDS) in phosphate
buffer (P+SDS), (3) 8 M urea in phosphate buffer (P+U), (4) 0.1 M 2-mercaptoethanol (2ME) in phosphate buffer (P+M), (5) 1.5% SDS and 8 M urea in phosphate buffer

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(P+S+U), (6) 1.5% SDS and 0.1 M 2-ME in phosphate buffer (P+S+M), (7) 8 M urea and
0.1 M 2-ME in phosphate buffer (P+U+M).
Approximately 200 g extrudate was finely chopped using a blender for 1 min. Sample (2
g) was extracted with 40 ml of each buffer for 2.5 h. in a 40C shaking water bath. It was
then centrifuged at 20,000 g for 40 min to obtain the supernatant. Protein concentration
was determined using Lowry method assay at 750 nm (UV/Visible 916
spectrophotometer, GBC Scientific Equipment Ltd., Australia).
Structural evaluation
The expansion ratio was defined as the rate of the diameter of extrudate over the diameter
of the die. The average for each data was obtained from twenty measurements. The
textural appearance of soy protein meat analog was determined using the modified sensory
method of Simonsky and Stanley (1982). The meat analog was cut in 5 cm lengths and
rehydrated for 5 min at room temperature. The rehydrated samples were then drained on a
screen. Quantitative descriptive analysis (QDA) was used as a technique to evaluate the
textural appearance of the meat analog using ten trained panelists. Individual strands of
meat analog were torn in a longitudinal axis by a judge to observe tearing attributes and
fibrous characteristics. A scale of 10 cm was used with two descriptors, tearing and
fibrousness. The replication of QDA was conducted in each experiment.
Textural measurement.
Meat analogs were cut in 3 cm lengths and rehydrated to 80% moisture at room
temperature. The texture of soy protein meat analog was measured using TA-XT2 Texture
Analyzer (Stable Micro Systems, UK) with WarnerBraztler shear cell. Cross head speed
was 2 mm/s. The maximum force and radius of each sample were recorded. Fifteen
measurements were conducted for each sample. The cutting force of extrudate (g/cm) was
reported as maximum force per cross-sectional area:
Statistical analysis
Completely randomized design (CRD) was the method employed and the data were
analyzed with the analysis of variance (ANOVA) and the average of each treatment was
compared using Duncans multiple range test (DMRT).
Results and Discussion
Effect of soy protein isolate on chemical linkages
Protein solubility was introduced to study the intermolecular chemical linkages of protein
structure. Urea, SDS and 2-ME were used to break the intramolecular H bond,
hydrophobic interaction and disulfide bonds between protein molecules respectively.
Disulfide bonds, hydrophobic interaction and hydrogen bonds were formed in the protein
structure of meat analog as shown in Table 1. Meat analog containing 20, 40, 60 and 80%
SPI showed the highest soluble protein in P+S+M and P+U+M, than that of other buffers
(p<0.05). In addition, soluble protein of other buffers was not significantly different

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(p>0.05). This result indicated that disulfide bonds, hydrophobic interaction and hydrogen
bonds were the major linkages in meat analog structure. It is in accordance with Hager
(1984) and PredencioFerrieira and Areas (1993) who reported the importance of disulfide
bonds, hydrophobic interaction and electrostatistic interaction in support of soy protein
extrudate structure.
Table 1. Soluble protein (g/g protein) of soy protein meat analog containing 20 to 80% in
7 buffers.

SPI content
/Buffer
20%SPI
40%SPI
60%SPI
80%SPI

P(1)

P+S(2)

P+U(3)

P+M(4) P+S+U(5) P+S+M(6) P+U+M(7)

0.0816y(8) 0.1687y

0.2949y

0.2462y

0.2085y

0.7994x

0.7198x

0.0817y

0.2107y

0.1723y

0.1749y

0.1705y

0.8367x

0.7952x

0.1172y

0.2816y

0.2406y

0.2512y

0.1352y

1.0184x

1.0122x

0.0540y

0.1369y

0.1668y

0.1886y

0.1051y

0.6307x

0.7117x

(1)

phosphate buffer
phosphate buffer containing sodium dodecyl sulphate
(3)
phosphate buffer containing urea
(4)
phosphate buffer containing 2-mercaptoethanol
(5)
phosphate buffer containing sodium dodecyl sulphate and urea
(6)
phosphate buffer containing sodium dodecyl sulphate and 2- mercaptoethanol
(7)
phosphate buffer containing urea and 2- mercaptoethanol
(8)
Different letters within the same row are significantly different (p<0.05).
(2)

The chemical linkages of material feed were not different from the meat analog including
disulfide bonds, hydrophobic interaction and hydrogen bonds as observed from the high
soluble protein in P+S, P+U, P+M and P+S+U of all materials and their extrudates
(p<0.05) (Table 2). Therefore, the extrusion did not alter the type of chemical bond
between material and meat analog. The study of SPI content on chemical bond formation
in meat analog indicated that increasing SPI content in meat analog did not affect the level
of chemical linkage cross-linked in protein molecules. An increase in SPI from 20 to 80%
in meat analog did not demonstrate different soluble protein in most of the buffers
(p>0.05). Thus, increasing protein content from 58 to 82% did not influence chemical
bonds in protein molecules of meat analog.

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Table 2. Soluble protein (g/g protein) of raw material containing 20 to 80 % in 7 buffers.

SPI content
/Buffer
20%SPI
40%SPI
60%SPI
80%SPI
(1)

P+S

P+U

P+M

P+S+U

P+S+M P+U+M

0.4542c(1) 0.7473abc

0.7168abc 0.5621bc

0.7466abc

0.9888a

0.8932 ab

0.4581c

0.8128a

0.7549a

0.6780ab

0.6406 ab

0.8776a

0.8982

0.4472 b

0.7695ab

0.8467ab 0.6875ab

0.6276ab

1.0058 a

0.9503a

0.3255b

0.6231ab

0.7006ab 0.5900ab

0.4431 ab

0.8664a

0.7974 a

Different letters within the row are significantly different (p<0.05).

Meat analog containing 20, 40, 60 and 80% SPI showed lower soluble protein content
than that of feed containing the same SPI content which was extracted by P, P+S, P+U,
P+M, P+S+U (p<0.05). This may result from protein denaturation by heat and pressure
during the extrusion process and protein reoriented to form the more complex structure. It
decreased protein solubility of the extrudate (Jeunink and Cheftel,1979; Noguchi et al.,
1981). Lin et al. (2000) reported that the high molecular weights of soy protein molecules
were formed by extrusion, resulting in the lower solubility of protein molecules. This is in
accord with the result of lower protein solubility of meat analogs as compared with its
feed material.
Effect of soy protein on physical characteristics
Increasing SPI content from 20 to 80% showed that the expansion ratio of meat analog
was significantly increased from 2.01 to 2.61 (Table 3). It indicated that material
components had an influence on the expansion ratio of meat analog. Yuryev et al., (1995)
reported that the damage of dispersed phase from shear stress during flowing in the
extruder affected viscosity and elasticity of the system which had an impact on expansion
ratio of the product.
Tolstoguzov et al., (1985) and Tolstoguzov (1993) reported that during extrusion
molecular weights of biopolymer occurred and incorporated to form an incompatible
structure leading to multiple phases being separated. The major component became a
continuous phase which was the main structure of the product. The other components
became a continuous phase which imparted overall characteristics of the product. A high
expansion appeared in high SPI meat analog, indicating that SPI provided elasticity to
meat analog dough, but less viscosity was found as indicated by a significant decrease
(P<0.05) in torque and die pressure of meat analog containing 80% SPI. The dough greatly
expanded after exiting from the die. The expansion of extrudate was a result of the
superheated steam flashing when the plastic-like dough exited from the die (Harper,
1981). This result was in accord with Rhee et al. (1981) who reported that meat analog

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containing higher SPI content had a lower specific density. Bhattacharya and Hanna
(1988) also found that high soy protein concentrate added into corn gluten resulted in
higher expansion of extrudate. On the contrary, meat analog containing low SPI showed
lower expansion ratio because it contained low protein and the dispersed melt
carbohydrate embedded on the protein matrix. Thus, the stretchability of protein
molecules was decreased resulting in the decreased expansion ratio of meat analog.
Table 3. Extrusion parameters and physical characteristics of meat analog containing 20
80% SPI.

Extrusion parameters
and characteristics
Torque (%)
Die pressure (psi)
Expansion ratio
Cutting force (g/cm2)
Textural appearance score
- Tearing
- Fibrousness
(1)

20
30.00ab(1)
183.33a
2.01d
2,477a
6.60a
5.36a

Soy protein isolate (%)

40
60

80

36.00a
193.33a
2.25c
2,186ab

35.00ab
196.67 a
2.36b
2,035 b

28.33 b
116.67 b
2.62 a
1,446 c

4.55b
3.85b

2.03 c
1.98 c

1.38 c
1.30 c

Different letters within the row are significant different (p<0.05).

The effect of SPI content on textural formation of meat analog was studied by the
measurement of textural cutting force. Table 3 shows that the meat analog containing 20%
SPI exhibited the maximum cutting force of 2,477 g/cm2 (P<0.05). Increasing SPI level
from 20 to 80% resulted in a reduction of cutting force from 2,477 to 1,446 g/cm2. It
indicated that the low SPI meat analog (58% protein) had a stronger layered structure than
that of higher SPI content (60, 74 and 82% protein). It could be explained that extrudate
with high SPI content had high expansion (Table 3) which contained large air cells of
loose structure. The low SPI meat analog comprised small air cells because there was low
protein to provide elasticity as well as carbohydrate embedded on the protein network:
thus, there was limited ability of mass dough to expand. This resulted in a stronger texture
of meat analog containing low SPI. This is also relevant to the results of Sheard et al.,
(1984) in that DSF extrudate showed a higher strength when compared to SPI extrudate.
The structural appearance characteristics of meat analog were evaluated using QDA
technique as shown in Table 3. The aim of this study was to develop the fibrous structure
of the soy protein meat analog which resemble meat muscle fibres. Therefore, the

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characteristics of tearing along the length of extrudate and fibre or layer structure of meat
analog were the main parameters of appearance structure examined. Meat analog
containing 20% SPI showed the highest tearing and fibrous structure (p<0.05) (Table3).
An increase in SPI level decreased the tearing and fibrousness of extruded product
(p<0.05). This was probably due to the higher SPI meat analog showing a higher
expansion ratio.
The lowest torque and die pressure was found in extrudate containing 80% SPI. It was not
able to build up enough pressure to form the finely fibrous structure during extrusion as
supported by Lin et al., (2002). Torque and die pressure of dough were the extrusion
parameter variables which were influenced by dough viscosity. An increase in dough
viscosity elevated torque on the screw drive and resulted in a high die pressure. Thus, high
SPI meat analog dough showed low viscosity, torque and die pressure which resulted in a
reduction of fibrous structure of meat analog.
Conclusion
The protein molecule in the raw material of meat analog was disrupted by extrusion
process. The chemical linkages within protein molecules were broken down together with
protein arrangement. This lead to the textural and structural formation of meat analog.
Disulfide bonds, hydrophobic interaction and hydrogen bonds were formed to support the
product structure. Increasing SPI content from 20 to 80% did not influence the ratio of
these chemical linkages in meat analog but the expansion was greater. The cutting force,
tearing and fibrousness of high SPI meat analog were also lower. Meat analog containing
low SPI showed strong, continuous, long and finely fibrous structure.
References
1.

Bhattacharya, M., and Hanna, M.A. (1988). Effect of lipid on the properties of
extruded product. J. Food Sci. 53: 1230-1231.

2.

Burgess, L.D., and Stanley, D.W. (1976). A possible mechanism for thermal
texturization of soybean protein. J. Ints. Can. Sci. Technol. Aliment. 9(4):
228-229.

3.

deMan, F.M. (1990). Principles of food chemistry (2 nd ed). Van Nostrand

Reinhold. New York, USA.

4.

Ha, T.T. (1995). Texturization of low-fat extruded/expelled soy flour by twinscrew extruder. M.S. Thesis. University of Illinois, Urbana, USA. 200 p.

5.

Hager, D.F. (1984). Effects of extrusion upon soy concentrate solubility. J. Agric.
Food Chem. 32: 293-296.

6.

Harper, J.M. (1981). Extrusion of foods II. CRC Press. Florida, USA. p. 174.

7.

Hayakawa, I. (1992). Food processing by ultra high pressure twin screw extrusion.
Technomic Publishing . Pennsylvania, USA. p.153.

As. J. Food Ag-Ind. 2008, 1(02)

104

8.

Jeunink, J. and Cheftel, J. C. (1979). Chemical and physicochemical change in

field bean and soybean texturized by extrusion. J. Food Sci. 44: 1322-1328.

9.

Lin, S., Huff, H.E., and Hsieh, F. (2000). Texture and chemical characteristic of
soy protein meat analog extruded at high moisture. J. Agric. Food Chem.
65(2): 264-269.

10.

Lin, S., Huff, H.E., and Hsieh, F. (2002). Extrusion process parameters, sensory
characteristics, and structural properties of high moisture soy protein meat
analog. J. Food Sci. 67(3): 1066-1072.

11.

Liu, K. (1997). Soybeans; chemistry, technology and utilization. Chapman and

Hall. New York, USA. p.477.

12.

Maurice, T.J. and Stanley, D. W., (1978). Texture-Structure relationships in

texturized soy protein IV. Influence of process variables on extrusion texturization.
J. Ints. Can. Sci. Technol. Aliment. 11: 1-6.

13.

Noguchi, A. (1981). Extrusion cooking of high-moisture protein foods. In C.

Mercier, P. Linko, and J.M. Harper (eds.). Extrusion cooking. p. 343-370.
American Association of Cereal Chemists: St. Paul, Minn., USA.

14.

Predencio-Ferriera, S.H. and Areas, J. A. G. (1993). Protein-protein interaction in

the extrusion of soya at various temperature and moisture contents. J. of Food Sci.
58: 378-381.

15.

Rhee, K.C., Kuo, C. K., and Lusas, E. W. (1981). Texturization. In J.P. Cherry
(ed.). Protein functionality in food. American Chemistry Society. Washington DC.,
USA.

16.

Sheard, P.R., Ledward, D.A., and Mitchell, J.R. (1984). Role of carbohydrates in
soya extrusion. J. Food Tech. 19: 475-483.

17.

Simonsky, R. W. and Stanley, D.W. (1982). Texture-structure relationships in

textureized soy protein V. Influence of pH and protein acylation of extrusion
textureization. Can. Inst. Food Sci. Technol. J. 15(4): 294-301.

18.

Tolstoguzov, V.B., Gringerg, V.Y. and Gurov, A. N. Some physicochemical

approaches to the problem of protein texturization. J. of Agric. Food Chem.
32(2): 151-159.

19.

Tolstoguzov, V.B. (1993). Thermoplastic extrusion - the mechanism of the

formation of extrudate structure and properties. JAOCS. 7: 417-424.

20.

Yuryev, V.P., Zasypkin, D.V., Alexeyev, V.V. and Bogatyrye, V.A. N. (1995).
Expansion ratio of extrudates preparaed from potato starch-soybean protein
mixtures. Carbo Polymer. 26: 215-218.